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Performance of Copan WASP for Routine Urine Microbiology Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2016 Performance of copan WASP for routine urine microbiology Quiblier, Chantal ; Jetter, Marion ; Rominski, Mark ; Mouttet, Forouhar ; Böttger, Erik C ; Keller, Peter M ; Hombach, Michael Abstract: This study compared manual workup of urine clinical samples with fully automated WASPLab processing. As a first step two different inocula (1 and 10 ฀l) and different streaking patterns were compared using WASP and InoqulA BT™ instrumentation. A 10 ฀l inoculum produced significantly more single colonies than a 1 ฀l inoculum and automated streaking yielded significantly more single colonies as compared to manual streaking on whole plates (p<0.001). In a second step, 379 clinical urine samples were evaluated using WASP and manual workup. Average numbers of detected morphologies, recovered species, and CFU/ml of all 379 urine samples showed excellent agreement of WASPLab and manual workup. The percentage of clinical categorization of urine samples as ”positive” or ”negative” did not differ between automated and manual work-flow but within the positive samples automated processing by WASPLab resulted in the detection of more potential pathogens. In summary, the present study demonstrates that i) the streaking pattern, i.e. primarily the number of zigzags/length of streaking lines, is critical for optimizing the number of single colonies yielded from primary cultures of urine samples, ii) automated streaking by the WASP instrument was superior to manual streaking regarding the number of single colonies yielded, (for 32.2%) iii) automated streaking leads to higher numbers of detected morphologies (for 47.5%), species (for 17.4%) and pathogens (for 3.4%). The results of this study point to an improved quality of microbiological analyses and laboratory reports when using automated sample processing by WASP and WASPLab. DOI: https://doi.org/10.1128/JCM.02577-15 Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-120648 Journal Article Accepted Version Originally published at: Quiblier, Chantal; Jetter, Marion; Rominski, Mark; Mouttet, Forouhar; Böttger, Erik C; Keller, Peter M; Hombach, Michael (2016). Performance of copan WASP for routine urine microbiology. Journal of Clinical Microbiology, 54(3):585-592. DOI: https://doi.org/10.1128/JCM.02577-15 JCM Accepted Manuscript Posted Online 16 December 2015 J. Clin. Microbiol. doi:10.1128/JCM.02577-15 Copyright © 2015, American Society for Microbiology. All Rights Reserved. 1 Performance of Copan WASP for Routine Urine Microbiology 2 3 Authors: 4 Chantal Quiblier,a* Marion Jetter,a* Mark Rominski,a Forouhar Mouttet,a Erik C. Böttger,a 5 Peter M. Keller,a° Michael Hombacha°+ 6 7 * Both authors contributed equally as first authors. 8 ° Both authors contributed equally as last authors. 9 10 Authors‘ affiliation: 11 a Universität Zürich, Institut für Medizinische Mikrobiologie, Gloriastrasse 30/32, CH- 12 8006 Zürich, Switzerland. 13 14 + Corresponding Author: 15 Dr. Michael Hombach, Universität Zürich, Institut für Medizinische Mikrobiologie, 16 Gloriastrasse 30/32, CH-8006 Zürich, Switzerland, email: [email protected], 17 phone: +41 44 6342700. 18 19 Word Count: 3144 20 21 Keywords: Automated microbiology, urine, Copan, WASP. 22 Abstract 23 This study compared manual workup of urine clinical samples with fully automated 24 WASPLab processing. As a first step two different inocula (1 and 10 μl) and different 25 streaking patterns were compared using WASP and InoqulA BTTM instrumentation. A 10 26 μl inoculum produced significantly more single colonies than a 1 μl inoculum and 27 automated streaking yielded significantly more single colonies as compared to manual 28 streaking on whole plates (p<0.001). In a second step, 379 clinical urine samples were 29 evaluated using WASP and manual workup. Average numbers of detected 30 morphologies, recovered species, and CFU/ml of all 379 urine samples showed 31 excellent agreement of WASPLab and manual workup. The percentage of clinical 32 categorization of urine samples as “positive” or “negative” did not differ between 33 automated and manual work-flow but within the positive samples automated processing 34 by WASPLab resulted in the detection of more potential pathogens. In summary, the 35 present study demonstrates that i) the streaking pattern, i.e. primarily the number of 36 zigzags/length of streaking lines, is critical for optimizing the number of single colonies 37 yielded from primary cultures of urine samples, ii) automated streaking by the WASP 38 instrument was superior to manual streaking regarding the number of single colonies 39 yielded, (for 32.2%) iii) automated streaking leads to higher numbers of detected 40 morphologies (for 47.5%), species (for 17.4%) and pathogens (for 3.4%). The results of 41 this study point to an improved quality of microbiological analyses and laboratory 42 reports when using automated sample processing by WASP and WASPLab. Evaluation of Copan WASP for Urine Microbiology 2 43 Introduction 44 In recent years, clinical microbiology has been faced with dramatic changes, as 45 full laboratory automation (FLA) has started to enter diagnostic laboratories. This trend 46 to automation will affect to various extents economic efficiency, standardization, and 47 time-to-result of laboratory procedures (16). To date, clinical microbiology is still 48 predominately based on manual sample processing. Compared to clinical chemistry, 49 microbiological specimens display a significantly higher degree of complexity (2). Thus, 50 for many years the general perception was that clinical microbiology would be far too 51 complex to allow for automated processing, and that robots would not be able to replace 52 human operators. However, it has been demonstrated that automated inoculation of 53 samples can be superior to manual processing and that automated reading of disk 54 diffusion agar plates significantly increases precision of results (4, 5, 9, 13). Thus, 55 automated sample processing promises an improved standardization of sample 56 processing, incubation times, and plate reading protocols. 57 FLA systems have been developed by several companies. Currently, there are 58 two FLA solutions available, i.e. BD Kiestra total laboratory automation (TLATM; BD 59 Kiestra B.V., Drachten, Netherlands) and WASPLab (Copan Italia S.p.A., Brescia, Italy). 60 These systems use robotic systems for handling specimen containers and primary 61 culture inoculation specifically the Kiestra InoqulA and the Copan WASP. The 62 inoculated media are moved to automated incubators by conveyor belts with integrated 63 camera systems to capture plate images at given time points which is henceforth 64 referred to in this article as WASPLab. Currently, digital plate reading (DPR) is still 65 depending on highly skilled technologists, who can read plates “virtually” without Evaluation of Copan WASP for Urine Microbiology 3 66 physical interaction (15). First studies on FLA solutions suggest that productivity 67 indicators can be improved and diagnostic processes can be accelerated (3, 6, 7, 14). 68 In the present study, we compared workflows for urine sample processing using 69 the fully automated WASP and WASPLab systems and manual standard procedures. 70 Inoculation procedures critically influence further procedures such as specimen 71 identification (ID) and antimicrobial susceptibility testing (AST), mainly by the ability to 72 generate single colonies suitable for further processing. Therefore, both streaking 73 pattern and inoculation volume should be well evaluated prior to compare total manual 74 and automated workflows. Different guidelines exist for urine specimen inoculation, e.g. 75 U.S. and Canadian laboratories routinely plate 1 μl of urine specimens, whereas 76 European guidelines suggest 10 μl (5, 18). Therefore, this study was subdivided in two 77 parts: In part one, the ability to generate single colonies was compared for i) manual 78 and automated workflows and ii) for different automated streaking patterns using pure 79 and mixed cultures and different inocula (1 μl and 10 μl) in order to determine the 80 optimal streaking pattern. Part two of the present study compared total manual and 81 automated workflows regarding i) the number of detected CFU, ii) the number of 82 detected morphologies, iii) the number of recovered species, and iv) the number of 83 follow-up testing after initial plate reading, i.e. the number of identifications and 84 susceptibility tests, in 379 clinical urine samples. 85 86 Materials and Methods 87 Bacterial strains, clinical samples and growth conditions. Urine samples and 88 bacterial strains isolated from patient specimens used in this study were collected from Evaluation of Copan WASP for Urine Microbiology 4 89 January until April 2015 in the clinical laboratory of the Institute of Medical Microbiology, 90 University of Zurich. If not stated otherwise bacterial cultures were incubated at 35 °C 91 ±2 °C and 7.5% CO2 for 16h-20h. 92 Quantitative analysis of streaking pattern/single colony count. Manual, semi and 93 fully automated quantitative streaking patterns were evaluated by counting the number 94 of single colonies produced on whole plates. Manual streaking was always performed 95 by the same investigator with a 1 µl and a 10 µl calibrated plastic inoculation loop 96 (Copan Italia S.p.A., Brescia, Italy) resulting in 9-10
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